Human Cells and Tissues/Sheppard

项目总结(见说明)： 特发性肺纤维化(IPF)占间质性肺疾病的一半以上。 患有IPF的患者发展为进行性肺功能丧失，平均在确诊后3年死亡。肺泡11型细胞在IPF肺组织中发育不良。然而，它们在疾病发病机制中的确切作用仍不清楚。该项目研究了上皮细胞通过三个病理过程直接参与肺纤维化形成的新假说：内质网应激，上皮整合素avB6激活TGPB，上皮向间充质转化。考虑到这一点 假设，这项翻译计划项目拨款的主要目标是开发针对IPF肺泡11型细胞的这些病理过程的新药。 以支持方案项目赠款的总体目标。 人类组织核心将：1.为处理、分类和存储从IPF患者和正常对照组获得的人类生物样本提供集中资源；2.从正常和纤维化的人类肺获得高纯度的肺泡II型细胞分离株，用于测试候选药物化合物；3.为比较分析人类II型细胞、从正常肺和IPF肺分离的肺片段和切片中的蛋白质和基因表达提供集中资源。 核心将在诱导经历内质网应激、avB6介导的TGFb激活或EMT的细胞和组织上执行表达微阵列和miRNA阵列，在存在或不存在抑制这些途径的药物的情况下，识别可以由每个项目表征的机械信息生物标志物。来自这个集中资源的细胞和组织将被用于提炼针对三种上皮细胞依赖的促纤维化途径中的每一种的候选药物。项目1检查在IPF11型细胞和普通型11型细胞中操纵未折叠蛋白反应的化合物，IPF11型细胞表现出UPR增加，而普通型11型细胞通过衣霉素培养激活UPR。项目2将使用肺组织的碎片来测试通过整合素avB6抑制上皮细胞依赖的TGFb激活的化合物。对于项目3，人类组织核心的领导者开发了使用FACS分类来分离高纯度的新方法 正常人和纤维化人肺的肺泡型II型细胞群。这些细胞将用于一种新的EMT体外模型，以测试阻断TGPB信号转导和EMT的候选化合物。

PROJECT SUMMARY (See instructions): Idiopathic pulmonary fibrosis (IPF) accounts for more than half of the cases of interstitial lung disease. Patients suffering from IPF develop a progressive loss of lung function and die, on average, 3 years after diagnosis. Alveolar type 11 cells are abormal in IPF lungs. However, their precise role in disease pathogenesis remains undefined. This program project grant examines the novel hypothesis that epithelial cells directly participate in the formation of lung fibrosis via three pathologic processes: ER stress, activation of TGPB by the epithelial integrin avB6, and epithelial to mesenchymal transition. Considering this hypothesis, the primary objective of this translational program project grant centers on developing novel drugs that target these pathological processes in IPF alveolar type 11 cells. In support of the overall objectives of the program project grant. The Human tissue core will: 1. provide a centralized resource for processing, cataloging and storing human biological samples obtained from patients with IPF and normal controls, 2. obtain highly purified isolates of alveolar type II cells from normal and fibrotic human lungs for use in testing candidate drug compounds, and 3; provide a centralized resource forthe comparative analysis of protein and gene expression in human type II cells, lung fragments and slices isolated from normal and IPF lungs. The core will perform expression microarrays and miRNA arrays on cells and tissues induced to undergo ER stress, avB6-mediated TGFB activation or EMT in the presence or absence of drugs to inhibit these pathways, to identify mechanistically informative biomarkers that can be characterized by each project. The cells and tissues from this centralized resource will be used to refine candidate drugs targeting each of the three epithelial cell dependent profibrotic pathways. Project 1 to examine compounds manipulating the unfolded protein response both in IPF type 11 cells, which exhibit an increased UPR, and in normal type 11 cells whose UPR is activated by culture in tunicamycin. Fragments of lung tissue will be used in project 2 to test compounds inhibiting epithelial cell dependent TGFB activation by the integrin avB6. For project 3, the leader of the human tissue core developed novel methods using FACS sorting to isolate highly pure populations of alveolar type II cells from normal and fibrotic human lungs. These cells will be used in a novel in vitro model of EMT to test candidate compounds blocking TGPB signalling and EMT.